IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week10/Chemical and Light

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(Transformations in Shewie)
(Getting mtrB onto thermo and lac inducible systems)
Line 61: Line 61:
=Getting mtrB onto thermo and lac inducible systems=
=Getting mtrB onto thermo and lac inducible systems=
 +
 +
==Ligating mtrB TOPO with RBS==
 +
Grew up more mtrB TOPO for next week.
 +
 +
==PCR of mtrB with RBS + mtrB primers==
 +
Scheduled for next week.

Revision as of 01:25, 31 August 2008

Contents

Transformations in Shewie

About 250 ng of DNA added each time.

First Set 8/22

Plate Colonies Size Color
S1 P102 (P108+45) Kan5 colonies3mm to 5 mmPink
S1 P121+P38 (P124) 1:2.5 Kan A1 colony5mm to 7.5mmPink
S1 P28 (ColE1) Kan0 colonies
S1 P121+P38(P124) 1:2.5 Kan B0 colonies
S1 P121+P39 1:2.5 Dephos Kan A0 colonies
S1 Topo vector (puc) 2ul Kan X-gal>50 colonies1mmWhite
S1 Topo vector (puc) 4ul Kan X-gal>50 colonies1mmWhite
S1 P121+P39 1:2.5 Kan D0 colonies
S1 (-) ctrl (just cells) Kan0 colonies
S1 (+) ctrl (E1 p59b) Kan2 colonies5mm Pink

Poor transformation efficiency in the positive control suggests that there may be a problem with our cells. All samples, except for P102, were retransformed the following week with fresh cell cultures.

Second Set (with new cells) 8/27

Plate Marker Description
S1 P124 1:2.5 AKan14 2-3mm pink colonies, ~100 0.5mm white colonies
S1 P124 1:2.5 BKan2 pink 2mm colonies, >100 0.5mm white colonies
S1 P125 1:2.5 Dephos AKan2 pink 2mm colonies, >100 0.5mm white colonies
S1 P125 1:2.5 DKan8 2mm pink colonies, ~100 0.5mm white colonies
S1 P28Kan~8 1-2mm pink colonies, ~50 0.5mm white colonies
S1 TOPO 2uLKanLawn of (>200) 0.5mm white colonies
S1 TOPO 4 uL (w/ Xgal)KanLawn of (>200) 0.5mm white colonies
S1 P59b (+) ctrlKan~50 2mm pink colonies, ~50 0.5mm white colonies
S1 (-) ctrlKan50-100 0.5mm white colonies

Getting Thermoinducible and Lac Inducible GFP into pSC101

Getting mtrB onto thermo and lac inducible systems

Ligating mtrB TOPO with RBS

Grew up more mtrB TOPO for next week.

PCR of mtrB with RBS + mtrB primers

Scheduled for next week.

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